LOCUS V045263 9387 bp DNA circular SYN 01-JAN-1980 DEFINITION synthetic circular DNA. ACCESSION V045263 VERSION V045263 KEYWORDS . SOURCE . ORGANISM . . FEATURES Location/Qualifiers LTR 1..634 /note="3' long terminal repeat (LTR) from HIV-1" /label="3' LTR" misc_feature 681..806 /note="packaging signal of human immunodeficiency virus type 1" /label="HIV-1 Ψ" misc_feature 1303..1536 /note="The Rev response element (RRE) of HIV-1 allows for Rev-dependent mRNA export from the nucleus to the cytoplasm." /label="RRE" CDS 1721..1765 /codon_start=1 /note="recognized by the 2H10 single-chain llama nanobody" /product="antigenic peptide corresponding to amino acids 655 to 669 of the HIV envelope protein gp41 (Lutje Hulsik et al., 2013)" /transl_table=1 /translation="KNEQELLELDKWASL" /label="gp41 peptide" misc_feature 2028..2143 /note="central polypurine tract and central termination sequence of HIV-1 (lacking the first T)" /label="cPPT/CTS" enhancer 2201..2504 /note="human cytomegalovirus immediate early enhancer" /label="CMV enhancer" promoter 2505..2708 /note="human cytomegalovirus (CMV) immediate early promoter" /label="CMV promoter" CDS 2827..3411 /label="PYCARD(NM_013258)" /note="PYCARD(NM_013258)" /gene="PYCARD" CDS 3442..4158 /codon_start=1 /note="mammalian codon-optimized" /product="monomeric green fluorescent protein, also known as mTagGFP" /transl_table=1 /translation="MSGGEELFAGIVPVLIELDGDVHGHKFSVRGEGEGDADYGKLEIK FICTTGKLPVPWPTLVTTLCYGIQCFARYPEHMKMNDFFKSAMPEGYIQERTIQFQDDG KYKTRGEVKFEGDTLVNRIELKGKDFKEDGNILGHKLEYSFNSHNVYIRPDKANNGLEA NFKTRHNIEGGGVQLADHYQTNVPLGDGPVLIPINHYLSTQTKISKDRNEARDHMVLLE SFSACCHTHGMDELYR*" /label="TagGFP2" promoter 4175..4674 /note="mouse phosphoglycerate kinase 1 promoter" /label="PGK promoter" CDS 4695..5294 /codon_start=1 /gene="pac from Streptomyces alboniger" /note="confers resistance to puromycin" /product="puromycin N-acetyltransferase" /transl_table=1 /translation="MTEYKPTVRLATRDDVPRAVRTLAAAFADYPATRHTVDPDRHIER VTELQELFLTRVGLDIGKVWVADDGAAVAVWTTPESVEAGAVFAEIGPRMAELSGSRLA AQQQMEGLLAPHRPKEPAWFLATVGVSPDHQGKGLGSAVVLPGVEAAERAGVPAFLETS APRNLPFYERLGFTVTADVEVPEGPRTWCMTRKPGA*" /label="PuroR" misc_feature 5308..5896 /note="woodchuck hepatitis virus posttranscriptional regulatory element" /label="WPRE" LTR 6103..6736 /note="3' long terminal repeat (LTR) from HIV-1" /label="3' LTR" primer_bind complement(6865..6881) /note="common sequencing primer, one of multiple similar variants" /label="M13 rev" protein_bind 6889..6905 /bound_moiety="lac repressor encoded by lacI" /note="The lac repressor binds to the lac operator to inhibit transcription in E. coli. This inhibition can be relieved by adding lactose or isopropyl-β-D-thiogalactopyranoside (IPTG)." /label="lac operator" promoter complement(6913..6943) /note="promoter for the E. coli lac operon" /label="lac promoter" protein_bind 6958..6979 /bound_moiety="E. coli catabolite activator protein" /note="CAP binding activates transcription in the presence of cAMP." /label="CAP binding site" rep_origin complement(7267..7855) /direction=LEFT /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" /label="ori" CDS complement(8026..8886) /codon_start=1 /gene="bla" /note="confers resistance to ampicillin, carbenicillin, and related antibiotics" /product="β-lactamase" /transl_table=1 /translation="MSIQHFRVALIPFFAAFCLPVFA,HPETLVKVKDAEDQLGARVGY IELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEY SPVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDR WEPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRS ALPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGA SLIKHW*" /label="AmpR" promoter complement(8887..8991) /gene="bla" /label="AmpR promoter" polyA_signal 9039..9173 /note="SV40 polyadenylation signal" /label="SV40 poly(A) signal" ORIGIN 1 tggaagggct aattcactcc caaagaagac aagatatcct tgatctgtgg atctaccaca 61 cacaaggcta cttccctgat tagcagaact acacaccagg gccaggggtc agatatccac 121 tgacctttgg atggtgctac aagctagtac cagttgagcc agataaggta gaagaggcca 181 ataaaggaga gaacaccagc ttgttacacc ctgtgagcct gcatgggatg gatgacccgg 241 agagagaagt gttagagtgg aggtttgaca gccgcctagc atttcatcac gtggcccgag 301 agctgcatcc ggagtacttc aagaactgct gatatcgagc ttgctacaag ggactttccg 361 ctggggactt tccagggagg cgtggcctgg gcgggactgg ggagtggcga gccctcagat 421 cctgcatata agcagctgct ttttgcctgt actgggtctc tctggttaga ccagatctga 481 gcctgggagc tctctggcta actagggaac ccactgctta agcctcaata aagcttgcct 541 tgagtgcttc aagtagtgtg tgcccgtctg ttgtgtgact ctggtaacta gagatccctc 601 agaccctttt agtcagtgtg gaaaatctct agcagtggcg cccgaacagg gacttgaaag 661 cgaaagggaa accagaggag ctctctcgac gcaggactcg gcttgctgaa gcgcgcacgg 721 caagaggcga ggggcggcga ctggtgagta cgccaaaaat tttgactagc ggaggctaga 781 aggagagaga tgggtgcgag agcgtcagta ttaagcgggg gagaattaga tcgcgatggg 841 aaaaaattcg gttaaggcca gggggaaaga aaaaatataa attaaaacat atagtatggg 901 caagcaggga gctagaacga ttcgcagtta atcctggcct gttagaaaca tcagaaggct 961 gtagacaaat actgggacag ctacaaccat cccttcagac aggatcagaa gaacttagat 1021 cattatataa tacagtagca accctctatt gtgtgcatca aaggatagag ataaaagaca 1081 ccaaggaagc tttagacaag atagaggaag agcaaaacaa aagtaagacc accgcacagc 1141 aagcggccgg ccgctgatct tcagacctgg aggaggagat atgagggaca attggagaag 1201 tgaattatat aaatataaag tagtaaaaat tgaaccatta ggagtagcac ccaccaaggc 1261 aaagagaaga gtggtgcaga gagaaaaaag agcagtggga ataggagctt tgttccttgg 1321 gttcttggga gcagcaggaa gcactatggg cgcagcgtca atgacgctga cggtacaggc 1381 cagacaatta ttgtctggta tagtgcagca gcagaacaat ttgctgaggg ctattgaggc 1441 gcaacagcat ctgttgcaac tcacagtctg gggcatcaag cagctccagg caagaatcct 1501 ggctgtggaa agatacctaa aggatcaaca gctcctgggg atttggggtt gctctggaaa 1561 actcatttgc accactgctg tgccttggaa tgctagttgg agtaataaat ctctggaaca 1621 gatttggaat cacacgacct ggatggagtg ggacagagaa attaacaatt acacaagctt 1681 aatacactcc ttaattgaag aatcgcaaaa ccagcaagaa aagaatgaac aagaattatt 1741 ggaattagat aaatgggcaa gtttgtggaa ttggtttaac ataacaaatt ggctgtggta 1801 tataaaatta ttcataatga tagtaggagg cttggtaggt ttaagaatag tttttgctgt 1861 actttctata gtgaatagag ttaggcaggg atattcacca ttatcgtttc agacccacct 1921 cccaaccccg aggggacccg acaggcccga aggaatagaa gaagaaggtg gagagagaga 1981 cagagacaga tccattcgat tagtgaacgg atctcgacgg tatcgccttt aaaagaaaag 2041 gggggattgg ggggtacagt gcaggggaaa gaatagtaga cataatagca acagacatac 2101 aaactaaaga attacaaaaa caaattacaa aaattcaaaa ttttcgggtt tattacaggg 2161 acagcagaga tccagtttat cgataagctt gggagttccg cgttacataa cttacggtaa 2221 atggcccgcc tggctgaccg cccaacgacc cccgcccatt gacgtcaata atgacgtatg 2281 ttcccatagt aacgccaata gggactttcc attgacgtca atgggtggag tatttacggt 2341 aaactgccca cttggcagta catcaagtgt atcatatgcc aagtacgccc cctattgacg 2401 tcaatgacgg taaatggccc gcctggcatt atgcccagta catgacctta tgggactttc 2461 ctacttggca gtacatctac gtattagtca tcgctattac catggtgatg cggttttggc 2521 agtacatcaa tgggcgtgga tagcggtttg actcacgggg atttccaagt ctccacccca 2581 ttgacgtcaa tgggagtttg ttttggcacc aaaatcaacg ggactttcca aaatgtcgta 2641 acaactccgc cccattgacg caaatgggcg gtaggcgtgt acggtgggag gtctatataa 2701 gcagagctcg tttagtgaac cgtcagatcg cctggagacg ccatccacgc tgttttgacc 2761 tccatagaag acaccgactc tactagagga tcgctagcgc taccggactc agatctcgag 2821 gccaccatgg ggcgcgcgcg cgacgccatc ctggatgcgc tggagaacct gaccgccgag 2881 gagctcaaga agttcaagct gaagctgctg tcggtgccgc tgcgcgaggg ctacgggcgc 2941 atcccgcggg gcgcgctgct gtccatggac gccttggacc tcaccgacaa gctggtcagc 3001 ttctacctgg agacctacgg cgccgagctc accgctaacg tgctgcgcga catgggcctg 3061 caggagatgg ccgggcagct gcaggcggcc acgcaccagg gctctggagc cgcgccagct 3121 gggatccagg cccctcctca gtcggcagcc aagccaggcc tgcactttat agaccagcac 3181 cgggctgcgc ttatcgcgag ggtcacaaac gttgagtggc tgctggatgc tctgtacggg 3241 aaggtcctga cggatgagca gtaccaggca gtgcgggccg agcccaccaa cccaagcaag 3301 atgcggaagc tcttcagttt cacaccagcc tggaactgga cctgcaagga cttgctcctc 3361 caggccctaa gggagtccca gtcctacctg gtggaggacc tggagcggag cggaggagga 3421 ggatcaggag gaggaggatc catgagcggg ggcgaggagc tgttcgccgg catcgtgccc 3481 gtgctgatcg agctggacgg cgacgtgcac ggccacaagt tcagcgtgcg cggcgagggc 3541 gagggcgacg ccgactacgg caagctggag atcaagttca tctgcaccac cggcaagctg 3601 cccgtgccct ggcccaccct ggtgaccacc ctctgctacg gcatccagtg cttcgcccgc 3661 taccccgagc acatgaagat gaacgacttc ttcaagagcg ccatgcccga gggctacatc 3721 caggagcgca ccatccagtt ccaggacgac ggcaagtaca agacccgcgg cgaggtgaag 3781 ttcgagggcg acaccctggt gaaccgcatc gagctgaagg gcaaggactt caaggaggac 3841 ggcaacatcc tgggccacaa gctggagtac agcttcaaca gccacaacgt gtacatccgc 3901 cccgacaagg ccaacaacgg cctggaggct aacttcaaga cccgccacaa catcgagggc 3961 ggcggcgtgc agctggccga ccactaccag accaacgtgc ccctgggcga cggccccgtg 4021 ctgatcccca tcaaccacta cctgagcact cagaccaaga tcagcaagga ccgcaacgag 4081 gcccgcgacc acatggtgct cctggagtcc ttcagcgcct gctgccacac ccacggcatg 4141 gacgagctgt acaggtaatc tagataattc taccgggtag gggaggcgct tttcccaagg 4201 cagtctggag catgcgcttt agcagccccg ctgggcactt ggcgctacac aagtggcctc 4261 tggcctcgca cacattccac atccaccggt aggcgccaac cggctccgtt ctttggtggc 4321 cccttcgcgc caccttctac tcctccccta gtcaggaagt tcccccccgc cccgcagctc 4381 gcgtcgtgca ggacgtgaca aatggaagta gcacgtctca ctagtctcgt gcagatggac 4441 agcaccgctg agcaatggaa gcgggtaggc ctttggggca gcggccaata gcagctttgc 4501 tccttcgctt tctgggctca gaggctggga aggggtgggt ccgggggcgg gctcaggggc 4561 gggctcaggg gcggggcggg cgcccgaagg tcctccggag gcccggcatt ctgcacgctt 4621 caaaagcgca cgtctgccgc gctgttctcc tcttcctcat ctccgggcct ttcgacctgc 4681 agcccaagct taccatgacc gagtacaagc ccacggtgcg cctcgccacc cgcgacgacg 4741 tccccagggc cgtacgcacc ctcgccgccg cgttcgccga ctaccccgcc acgcgccaca 4801 ccgtcgatcc ggaccgccac atcgagcggg tcaccgagct gcaagaactc ttcctcacgc 4861 gcgtcgggct cgacatcggc aaggtgtggg tcgcggacga cggcgccgcg gtggcggtct 4921 ggaccacgcc ggagagcgtc gaagcggggg cggtgttcgc cgagatcggc ccgcgcatgg 4981 ccgagttgag cggttcccgg ctggccgcgc agcaacagat ggaaggcctc ctggcgccgc 5041 accggcccaa ggagcccgcg tggttcctgg ccaccgtcgg cgtctcgccc gaccaccagg 5101 gcaagggtct gggcagcgcc gtcgtgctcc ccggagtgga ggcggccgag cgcgccgggg 5161 tgcccgcctt cctggagacc tccgcgcccc gcaacctccc cttctacgag cggctcggct 5221 tcaccgtcac cgccgacgtc gaggtgcccg aaggaccgcg cacctggtgc atgacccgca 5281 agcccggtgc ctgaccgcgt ctggaacaat caacctctgg attacaaaat ttgtgaaaga 5341 ttgactggta ttcttaacta tgttgctcct tttacgctat gtggatacgc tgctttaatg 5401 cctttgtatc atgctattgc ttcccgtatg gctttcattt tctcctcctt gtataaatcc 5461 tggttgctgt ctctttatga ggagttgtgg cccgttgtca ggcaacgtgg cgtggtgtgc 5521 actgtgtttg ctgacgcaac ccccactggt tggggcattg ccaccacctg tcagctcctt 5581 tccgggactt tcgctttccc cctccctatt gccacggcgg aactcatcgc cgcctgcctt 5641 gcccgctgct ggacaggggc tcggctgttg ggcactgaca attccgtggt gttgtcgggg 5701 aagctgacgt cctttccatg gctgctcgcc tgtgttgcca cctggattct gcgcgggacg 5761 tccttctgct acgtcccttc ggccctcaat ccagcggacc ttccttcccg cggcctgctg 5821 ccggctctgc ggcctcttcc gcgtcttcgc cttcgccctc agacgagtcg gatctccctt 5881 tgggccgcct ccccgcctgg aattaattct gcagtcgaga cctagaaaaa catggagcaa 5941 tcacaagtag caatacagca gctaccaatg ctgattgtgc ctggctagaa gcacaagagg 6001 aggaggaggt gggttttcca gtcacacctc aggtaccttt aagaccaatg acttacaagg 6061 cagctgtaga tcttagccac tttttaaaag aaaagagggg actggaaggg ctaattcact 6121 cccaacgaag acaagatatc cttgatctgt ggatctacca cacacaaggc tacttccctg 6181 attagcagaa ctacacacca gggccagggg tcagatatcc actgaccttt ggatggtgct 6241 acaagctagt accagttgag ccagataagg tagaagaggc caataaagga gagaacacca 6301 gcttgttaca ccctgtgagc ctgcatggga tggatgaccc ggagagagaa gtgttagagt 6361 ggaggtttga cagccgccta gcatttcatc acgtggcccg agagctgcat ccggagtact 6421 tcaagaactg ctgatatcga gcttgctaca agggactttc cgctggggac tttccaggga 6481 ggcgtggcct gggcgggact ggggagtggc gagccctcag atcctgcata taagcagctg 6541 ctttttgcct gtactgggtc tctctggtta gaccagatct gagcctggga gctctctggc 6601 taactaggga acccactgct taagcctcaa taaagcttgc cttgagtgct tcaagtagtg 6661 tgtgcccgtc tgttgtgtga ctctggtaac tagagatccc tcagaccctt ttagtcagtg 6721 tggaaaatct ctagcagtag tagttcatgt catcttatta ttcagtattt ataacttgca 6781 aagaaatgaa tatcagagag tgagaggcct tgacattgct agcgttttac cgtcgacctc 6841 tagctagagc ttggcgtaat catggtcata gctgtttcct gtgtgaaatt gttatccgct 6901 cacaattcca cacaacatac gagccggaag cataaagtgt aaagcctggg gtgcctaatg 6961 agtgagctaa ctcacattaa ttgcgttgcg ctcactgccc gctttccagt cgggaaacct 7021 gtcgtgccag ctgcattaat gaatcggcca acgcgcgggg agaggcggtt tgcgtattgg 7081 gcgctcttcc gcttcctcgc tcactgactc gctgcgctcg gtcgttcggc tgcggcgagc 7141 ggtatcagct cactcaaagg cggtaatacg gttatccaca gaatcagggg ataacgcagg 7201 aaagaacatg tgagcaaaag gccagcaaaa ggccaggaac cgtaaaaagg ccgcgttgct 7261 ggcgtttttc cataggctcc gcccccctga cgagcatcac aaaaatcgac gctcaagtca 7321 gaggtggcga aacccgacag gactataaag ataccaggcg tttccccctg gaagctccct 7381 cgtgcgctct cctgttccga ccctgccgct taccggatac ctgtccgcct ttctcccttc 7441 gggaagcgtg gcgctttctc atagctcacg ctgtaggtat ctcagttcgg tgtaggtcgt 7501 tcgctccaag ctgggctgtg tgcacgaacc ccccgttcag cccgaccgct gcgccttatc 7561 cggtaactat cgtcttgagt ccaacccggt aagacacgac ttatcgccac tggcagcagc 7621 cactggtaac aggattagca gagcgaggta tgtaggcggt gctacagagt tcttgaagtg 7681 gtggcctaac tacggctaca ctagaagaac agtatttggt atctgcgctc tgctgaagcc 7741 agttaccttc ggaaaaagag ttggtagctc ttgatccggc aaacaaacca ccgctggtag 7801 cggtggtttt tttgtttgca agcagcagat tacgcgcaga aaaaaaggat ctcaagaaga 7861 tcctttgatc ttttctacgg ggtctgacgc tcagtggaac gaaaactcac gttaagggat 7921 tttggtcatg agattatcaa aaaggatctt cacctagatc cttttaaatt aaaaatgaag 7981 ttttaaatca atctaaagta tatatgagta aacttggtct gacagttacc aatgcttaat 8041 cagtgaggca cctatctcag cgatctgtct atttcgttca tccatagttg cctgactccc 8101 cgtcgtgtag ataactacga tacgggaggg cttaccatct ggccccagtg ctgcaatgat 8161 accgcgagac ccacgctcac cggctccaga tttatcagca ataaaccagc cagccggaag 8221 ggccgagcgc agaagtggtc ctgcaacttt atccgcctcc atccagtcta ttaattgttg 8281 ccgggaagct agagtaagta gttcgccagt taatagtttg cgcaacgttg ttgccattgc 8341 tacaggcatc gtggtgtcac gctcgtcgtt tggtatggct tcattcagct ccggttccca 8401 acgatcaagg cgagttacat gatcccccat gttgtgcaaa aaagcggtta gctccttcgg 8461 tcctccgatc gttgtcagaa gtaagttggc cgcagtgtta tcactcatgg ttatggcagc 8521 actgcataat tctcttactg tcatgccatc cgtaagatgc ttttctgtga ctggtgagta 8581 ctcaaccaag tcattctgag aatagtgtat gcggcgaccg agttgctctt gcccggcgtc 8641 aatacgggat aataccgcgc cacatagcag aactttaaaa gtgctcatca ttggaaaacg 8701 ttcttcgggg cgaaaactct caaggatctt accgctgttg agatccagtt cgatgtaacc 8761 cactcgtgca cccaactgat cttcagcatc ttttactttc accagcgttt ctgggtgagc 8821 aaaaacagga aggcaaaatg ccgcaaaaaa gggaataagg gcgacacgga aatgttgaat 8881 actcatactc ttcctttttc aatattattg aagcatttat cagggttatt gtctcatgag 8941 cggatacata tttgaatgta tttagaaaaa taaacaaata ggggttccgc gcacatttcc 9001 ccgaaaagtg ccacctgacg tcgacggatc gggagatcaa cttgtttatt gcagcttata 9061 atggttacaa ataaagcaat agcatcacaa atttcacaaa taaagcattt ttttcactgc 9121 attctagttg tggtttgtcc aaactcatca atgtatctta tcatgtctgg atcaactgga 9181 taactcaagc taaccaaaat catcccaaac ttcccacccc ataccctatt accactgcca 9241 attacctgtg gtttcattta ctctaaacct gtgattcctc tgaattattt tcattttaaa 9301 gaaattgtat ttgttaaata tgtactacaa acttagtagt ttttaaagaa attgtatttg 9361 ttaaatatgt actacaaact tagtagt //